Cargando…

Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa

BACKGROUND: The genus Planococcus is comprised of halophilic bacteria generally reported for the production of carotenoid pigments and biosurfactants. In previous work, we showed that the culturing of the orange-pigmented Planococcus sp. CP5-4 isolate increased the evaporation rate of industrial was...

Descripción completa

Detalles Bibliográficos
Autores principales: Moyo, Anesu Conrad, Dufossé, Laurent, Giuffrida, Daniele, van Zyl, Leonardo Joaquim, Trindade, Marla
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8933910/
https://www.ncbi.nlm.nih.gov/pubmed/35305628
http://dx.doi.org/10.1186/s12934-022-01752-1
_version_ 1784671759390212096
author Moyo, Anesu Conrad
Dufossé, Laurent
Giuffrida, Daniele
van Zyl, Leonardo Joaquim
Trindade, Marla
author_facet Moyo, Anesu Conrad
Dufossé, Laurent
Giuffrida, Daniele
van Zyl, Leonardo Joaquim
Trindade, Marla
author_sort Moyo, Anesu Conrad
collection PubMed
description BACKGROUND: The genus Planococcus is comprised of halophilic bacteria generally reported for the production of carotenoid pigments and biosurfactants. In previous work, we showed that the culturing of the orange-pigmented Planococcus sp. CP5-4 isolate increased the evaporation rate of industrial wastewater brine effluent, which we attributed to the orange pigment. This demonstrated the potential application of this bacterium for industrial brine effluent management in evaporation ponds for inland desalination plants. Here we identified a C(30)-carotenoid biosynthetic gene cluster responsible for pigment biosynthesis in Planococcus sp. CP5-4 through isolation of mutants and genome sequencing. We further compare the core genes of the carotenoid biosynthetic gene clusters identified from different Planococcus species’ genomes which grouped into gene cluster families containing BGCs linked to different carotenoid product chemotypes. Lastly, LC–MS analysis of saponified and unsaponified pigment extracts obtained from cultures of Planococcus sp. CP5-4, revealed the structure of the main (predominant) glucosylated C(30)-carotenoid fatty acid ester produced by Planococcus sp. CP5-4. RESULTS: Genome sequence comparisons of isolated mutant strains of Planococcus sp. CP5-4 showed deletions of 146 Kb and 3 Kb for the non-pigmented and “yellow” mutants respectively. Eight candidate genes, likely responsible for C(30)-carotenoid biosynthesis, were identified on the wild-type genome region corresponding to the deleted segment in the non-pigmented mutant. Six of the eight candidate genes formed a biosynthetic gene cluster. A truncation of crtP was responsible for the “yellow” mutant phenotype. Genome annotation revealed that the genes encoded 4,4′-diapolycopene oxygenase (CrtNb), 4,4′- diapolycopen-4-al dehydrogenase (CrtNc), 4,4′-diapophytoene desaturase (CrtN), 4,4′- diaponeurosporene oxygenase (CrtP), glycerol acyltransferase (Agpat), family 2 glucosyl transferase 2 (Gtf2), phytoene/squalene synthase (CrtM), and cytochrome P450 hydroxylase enzymes. Carotenoid analysis showed that a glucosylated C(30)-carotenoid fatty acid ester, methyl 5-(6-C(17:3))-glucosyl-5, 6′-dihydro-apo-4, 4′-lycopenoate was the main carotenoid compound produced by Planococcus sp. CP5-4. CONCLUSION: We identified and characterized the carotenoid biosynthetic gene cluster and the C(30)-carotenoid compound produced by Planococcus sp. CP5-4. Mass-spectrometry guided analysis of the saponified and unsaponified pigment extracts showed that methyl 5-glucosyl-5, 6-dihydro-apo-4, 4′-lycopenoate esterified to heptadecatrienoic acid (C(17:3)). Furthermore, through phylogenetic analysis of the core carotenoid BGCs of Planococcus species we show that various C(30)-carotenoid product chemotypes, apart from methyl 5-glucosyl-5, 6-dihydro-apo-4, 4′-lycopenoate and 5-glucosyl-4, 4-diaponeurosporen-4′-ol-4-oic acid, may be produced that could offer opportunities for a variety of applications. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01752-1.
format Online
Article
Text
id pubmed-8933910
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-89339102022-03-23 Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa Moyo, Anesu Conrad Dufossé, Laurent Giuffrida, Daniele van Zyl, Leonardo Joaquim Trindade, Marla Microb Cell Fact Research BACKGROUND: The genus Planococcus is comprised of halophilic bacteria generally reported for the production of carotenoid pigments and biosurfactants. In previous work, we showed that the culturing of the orange-pigmented Planococcus sp. CP5-4 isolate increased the evaporation rate of industrial wastewater brine effluent, which we attributed to the orange pigment. This demonstrated the potential application of this bacterium for industrial brine effluent management in evaporation ponds for inland desalination plants. Here we identified a C(30)-carotenoid biosynthetic gene cluster responsible for pigment biosynthesis in Planococcus sp. CP5-4 through isolation of mutants and genome sequencing. We further compare the core genes of the carotenoid biosynthetic gene clusters identified from different Planococcus species’ genomes which grouped into gene cluster families containing BGCs linked to different carotenoid product chemotypes. Lastly, LC–MS analysis of saponified and unsaponified pigment extracts obtained from cultures of Planococcus sp. CP5-4, revealed the structure of the main (predominant) glucosylated C(30)-carotenoid fatty acid ester produced by Planococcus sp. CP5-4. RESULTS: Genome sequence comparisons of isolated mutant strains of Planococcus sp. CP5-4 showed deletions of 146 Kb and 3 Kb for the non-pigmented and “yellow” mutants respectively. Eight candidate genes, likely responsible for C(30)-carotenoid biosynthesis, were identified on the wild-type genome region corresponding to the deleted segment in the non-pigmented mutant. Six of the eight candidate genes formed a biosynthetic gene cluster. A truncation of crtP was responsible for the “yellow” mutant phenotype. Genome annotation revealed that the genes encoded 4,4′-diapolycopene oxygenase (CrtNb), 4,4′- diapolycopen-4-al dehydrogenase (CrtNc), 4,4′-diapophytoene desaturase (CrtN), 4,4′- diaponeurosporene oxygenase (CrtP), glycerol acyltransferase (Agpat), family 2 glucosyl transferase 2 (Gtf2), phytoene/squalene synthase (CrtM), and cytochrome P450 hydroxylase enzymes. Carotenoid analysis showed that a glucosylated C(30)-carotenoid fatty acid ester, methyl 5-(6-C(17:3))-glucosyl-5, 6′-dihydro-apo-4, 4′-lycopenoate was the main carotenoid compound produced by Planococcus sp. CP5-4. CONCLUSION: We identified and characterized the carotenoid biosynthetic gene cluster and the C(30)-carotenoid compound produced by Planococcus sp. CP5-4. Mass-spectrometry guided analysis of the saponified and unsaponified pigment extracts showed that methyl 5-glucosyl-5, 6-dihydro-apo-4, 4′-lycopenoate esterified to heptadecatrienoic acid (C(17:3)). Furthermore, through phylogenetic analysis of the core carotenoid BGCs of Planococcus species we show that various C(30)-carotenoid product chemotypes, apart from methyl 5-glucosyl-5, 6-dihydro-apo-4, 4′-lycopenoate and 5-glucosyl-4, 4-diaponeurosporen-4′-ol-4-oic acid, may be produced that could offer opportunities for a variety of applications. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01752-1. BioMed Central 2022-03-19 /pmc/articles/PMC8933910/ /pubmed/35305628 http://dx.doi.org/10.1186/s12934-022-01752-1 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Moyo, Anesu Conrad
Dufossé, Laurent
Giuffrida, Daniele
van Zyl, Leonardo Joaquim
Trindade, Marla
Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa
title Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa
title_full Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa
title_fullStr Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa
title_full_unstemmed Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa
title_short Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa
title_sort structure and biosynthesis of carotenoids produced by a novel planococcus sp. isolated from south africa
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8933910/
https://www.ncbi.nlm.nih.gov/pubmed/35305628
http://dx.doi.org/10.1186/s12934-022-01752-1
work_keys_str_mv AT moyoanesuconrad structureandbiosynthesisofcarotenoidsproducedbyanovelplanococcusspisolatedfromsouthafrica
AT dufosselaurent structureandbiosynthesisofcarotenoidsproducedbyanovelplanococcusspisolatedfromsouthafrica
AT giuffridadaniele structureandbiosynthesisofcarotenoidsproducedbyanovelplanococcusspisolatedfromsouthafrica
AT vanzylleonardojoaquim structureandbiosynthesisofcarotenoidsproducedbyanovelplanococcusspisolatedfromsouthafrica
AT trindademarla structureandbiosynthesisofcarotenoidsproducedbyanovelplanococcusspisolatedfromsouthafrica